GW501516 (GW-501516, MW 471.52 g/mol, CAS 317318-70-0) is a selective peroxisome proliferator-activated receptor delta (PPAR-delta/PPAR-beta) agonist studied in cell-based nuclear receptor pharmacology and metabolic pathway research. PPAR-delta is a ligand-activated transcription factor expressed in skeletal muscle, adipose, and liver cell lines.

Molecular Properties

| Property | Value |

|----------|--------|

| Molecular Formula | C₂₁H₁₈F₃NO₃S₂ |

| Molecular Weight | 471.52 g/mol |

| CAS Number | 317318-70-0 |

| Target | PPAR-delta (PPARbeta) |

PPAR-delta Receptor Pharmacology

Nuclear Receptor Structure and Function

PPAR-delta belongs to the nuclear hormone receptor superfamily, functioning as a ligand-dependent transcription factor. The receptor contains distinct functional domains including the DNA-binding domain (DBD) and ligand-binding domain (LBD). Upon ligand binding, PPAR-delta undergoes conformational changes that facilitate heterodimerization with retinoid X receptor (RXR), enabling recognition of specific DNA sequences called peroxisome proliferator response elements (PPREs).

Ligand-Binding Kinetics

GW501516 demonstrates high selectivity for PPAR-delta with reported binding affinity values in the nanomolar range. Radioligand binding assays using tritiated GW501516 reveal competitive binding kinetics with rapid association and relatively slow dissociation rates. The compound exhibits over 1000-fold selectivity for PPAR-delta compared to PPAR-alpha and PPAR-gamma subtypes, making it valuable for studying subtype-specific receptor pharmacology.

In Vitro Assay Applications

Cell-Based Reporter Systems

Transfection studies utilizing PPAR-delta responsive reporter constructs demonstrate dose-dependent transcriptional activation by GW501516. These luciferase-based assay systems typically employ GAL4-PPAR-delta chimeric receptors or native PPRE-driven reporter plasmids in cell lines such as CV-1 or HEK293. EC50 values for transcriptional activation generally fall within the 10-100 nanomolar range, correlating with binding affinity measurements.

Gene Expression Analysis

Real-time PCR analysis of PPAR-delta target genes provides insight into downstream transcriptional effects. GW501516 treatment upregulates expression of genes involved in fatty acid oxidation pathways, including carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACOX1), and uncoupling protein 3 (UCP3) in appropriate cell model systems.

Metabolic Pathway Studies

Fatty Acid Oxidation Enzymes

PPAR-delta activation by GW501516 modulates expression of key enzymes in beta-oxidation pathways. Cell culture studies demonstrate increased mRNA and protein levels of medium-chain acyl-CoA dehydrogenase (MCAD) and long-chain acyl-CoA dehydrogenase (LCAD). These enzymatic changes can be quantified through activity assays measuring substrate conversion rates.

Glucose Metabolism Pathways

Research using hepatocyte cell lines reveals GW501516 effects on gluconeogenic enzyme expression. The compound influences phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) expression levels, providing models for studying glucose production pathway regulation.

Signaling Pathway Interactions

AMPK Pathway Crosstalk

Studies indicate potential interactions between PPAR-delta signaling and AMP-activated protein kinase (AMPK) pathways. GW501516 treatment in cell culture models affects AMPK phosphorylation status and downstream target activation, suggesting coordinated metabolic regulation mechanisms.

Mitochondrial Biogenesis

PPAR-delta agonism influences mitochondrial biogenesis markers including peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1alpha) expression. Cell-based assays measuring mitochondrial DNA content and respiratory chain enzyme activities provide quantitative endpoints for studying these effects.

Research Applications

Primary Cell Culture Studies

GW501516 serves as a valuable tool compound in primary myotube, hepatocyte, and adipocyte culture systems for investigating PPAR-delta-mediated metabolic regulation. These models enable examination of native cellular responses without immortalized cell line artifacts.

Protein-Protein Interaction Studies

The compound facilitates studies of PPAR-delta coactivator and corepressor interactions using techniques such as mammalian two-hybrid systems, chromatin immunoprecipitation, and fluorescence polarization assays.

Research Summary

GW501516 represents a highly selective PPAR-delta agonist with well-characterized binding kinetics and transcriptional activation properties. Its utility in cell-based assay systems enables detailed investigation of nuclear receptor pharmacology, metabolic enzyme regulation, and signaling pathway interactions. The compound's selectivity profile and potent activity make it an essential research tool for studying PPAR-delta-mediated cellular processes in various in vitro model systems.

All content is intended for in vitro laboratory research purposes only. Not for human or animal consumption. Not intended to diagnose, treat, cure, or prevent any condition.